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Detection of residual rifampicin in urine via fluorescence quenching of gold nanoclusters on paper.

Chatterjee K, Kuo CW, Chen A, Chen P - J Nanobiotechnology (2015)

Bottom Line: The decreased fluorescence intensity of BSA-Au NCs in the presence of rifampicin allows for the sensitive detection of rifampicin in a range from 0.5 to 823 µg/mL.The detection limit for rifampicin was measured as 70 ng/mL.We have developed a robust, cost-effective, and portable point-of-care medical diagnostic platform for the detection of rifampicin in urine based on the ability of rifampicin to quench the fluorescence of immobilized BSA-Au NCs on wax-printed papers.

View Article: PubMed Central - PubMed

Affiliation: Department of Engineering and System Science, National Tsing Hua University, Hsinchu, 300, Taiwan. sanu.hit@gmail.com.

ABSTRACT

Background: Rifampicin or rifampin (R) is a common drug used to treat inactive meningitis, cholestatic pruritus and tuberculosis (TB), and it is generally prescribed for long-term administration under regulated dosages. Constant monitoring of rifampicin is important for controlling the side effects and preventing overdose caused by chronic medication. In this study, we present an easy to use, effective and less costly method for detecting residual rifampicin in urine samples using protein (bovine serum albumin, BSA)-stabilized gold nanoclusters (BSA-Au NCs) adsorbed on a paper substrate in which the concentration of rifampicin in urine can be detected via fluorescence quenching. The intensity of the colorimetric assay performed on the paper-based platforms can be easily captured using a digital camera and subsequently analyzed.

Results: The decreased fluorescence intensity of BSA-Au NCs in the presence of rifampicin allows for the sensitive detection of rifampicin in a range from 0.5 to 823 µg/mL. The detection limit for rifampicin was measured as 70 ng/mL. The BSA-Au NCs were immobilized on a wax-printed paper-based platform and used to conduct real-time monitoring of rifampicin in urine.

Conclusion: We have developed a robust, cost-effective, and portable point-of-care medical diagnostic platform for the detection of rifampicin in urine based on the ability of rifampicin to quench the fluorescence of immobilized BSA-Au NCs on wax-printed papers. The paper-based assay can be further used for the detection of other specific analytes via surface modification of the BSA in BSA-Au NCs and offers a useful tool for monitoring other diseases.

No MeSH data available.


Related in: MedlinePlus

a Fluorescence emission spectra of BSA-Au NCs (0.1× dilution) in the presence of primary TB drugs (100 µM each in the final concentration). At 640 nm (emission wavelength): BSA-Au NCs (fluorescence intensity = 123.74 ± 1.56), NCs + rifampicin (83 µg/mL) (13.67 ± 0.33), NCs + pyrazinamide (12.3 µg/mL) (121.95 ± 0.69), NCs + ethambutol (27.7 µg/mL) (114.27 ± 1.28) and NCs + izoniazid (13.7 µg/mL) (127 ± 0.69) (each data point represents the average of three separate studies (n = 3), and the error bars denote the standard error of measurements within each experiment). b Comparison of the normalized decrease in fluorescence intensity of BSA-Au NCs (0.1× dilution) in the presence of primary TB drugs (100 µM each in final concentration). At 640 nm (emission wavelength): normalized quenching of NCs + rifampicin [(F0 − F)/F0 ~ 0.89 ± 0.006], NCs + pyrazinamide [0.013 ± 0.005], NCs + ethambutol [0.065 ± 0.008] and NCs + izoniazid [0.028 ± 0.007] (each data point represents the average of three separate studies (n = 3), and the error bars denote the standard error of measurements within each experiment). [In this work, (F0 − F)/F0 = 1 indicates complete quenching and (F0 − F)/F0 = 0 indicates no quenching]. The excitation wavelength was set at 480 nm, and the emission wavelength was 640 nm.
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Fig4: a Fluorescence emission spectra of BSA-Au NCs (0.1× dilution) in the presence of primary TB drugs (100 µM each in the final concentration). At 640 nm (emission wavelength): BSA-Au NCs (fluorescence intensity = 123.74 ± 1.56), NCs + rifampicin (83 µg/mL) (13.67 ± 0.33), NCs + pyrazinamide (12.3 µg/mL) (121.95 ± 0.69), NCs + ethambutol (27.7 µg/mL) (114.27 ± 1.28) and NCs + izoniazid (13.7 µg/mL) (127 ± 0.69) (each data point represents the average of three separate studies (n = 3), and the error bars denote the standard error of measurements within each experiment). b Comparison of the normalized decrease in fluorescence intensity of BSA-Au NCs (0.1× dilution) in the presence of primary TB drugs (100 µM each in final concentration). At 640 nm (emission wavelength): normalized quenching of NCs + rifampicin [(F0 − F)/F0 ~ 0.89 ± 0.006], NCs + pyrazinamide [0.013 ± 0.005], NCs + ethambutol [0.065 ± 0.008] and NCs + izoniazid [0.028 ± 0.007] (each data point represents the average of three separate studies (n = 3), and the error bars denote the standard error of measurements within each experiment). [In this work, (F0 − F)/F0 = 1 indicates complete quenching and (F0 − F)/F0 = 0 indicates no quenching]. The excitation wavelength was set at 480 nm, and the emission wavelength was 640 nm.

Mentions: We tested the selectivity of BSA-Au NCs for sensing rifampicin in the presence of other antibiotics (INZ, E and P) used collaterally with rifampicin for the treatment of TB. The fluorescent responses of BSA-Au NCs with all four antibiotics were monitored and compared with that of rifampicin. Figure 4a shows that the addition of rifampicin at a final concentration of 100 µM (83 µg/mL) resulted in a nearly 90% decrease in the fluorescence intensity of the BSA-Au NCs, whereas a limited decrease in fluorescence intensity was observed for P (1.3%), INZ (2.8%) and E (6.5%) at the same molar concentration (Figure 4b). The clinical effectiveness of rifampicin as a primary drug for TB can mainly be attributed to 4-methyl-1-piperazinaminyl substitution on rifampicin. Generally, antibiotics with piperazine moiety exhibit good efficacy as therapeutic agents [42–44]. Therefore, we tested the specificity of BSA-Au NCs toward detecting rifampicin using antibiotics with piperazine functional group such as ciprofloxacin [42], buspirone [43], ipsapirone [44] (Additional file 1: Figure S10a). In addition, we also measured the fluorescence responses of some commonly used antibiotics and drugs (Additional file 1: Figure S10b). The average amount of residual rifampicin present in the patients’ urine is in the range of 45–55 µg/mL [45]. Our results showed that at a final concentration of 50 µg/mL of rifampicin a nearly 60% decrease in the fluorescence intensity of the BSA-Au NCs was measured, whereas only a limited decrease in fluorescence intensity (less than 10%, p < 0.001) was observed for all the other antibiotics and drugs at the same concentration. Therefore, BSA-Au NCs can be used for preferential sensing of rifampicin among other antibiotics administered at the same time. Additionally, the prepared BSA-Au NCs were freeze-dried and stored for 3 months before dispersion for the same experiment (Additional file 1: Figure S11). No drastic changes were observed, even after storage, indicating the excellent stability of the BSA-Au NC.Figure 4


Detection of residual rifampicin in urine via fluorescence quenching of gold nanoclusters on paper.

Chatterjee K, Kuo CW, Chen A, Chen P - J Nanobiotechnology (2015)

a Fluorescence emission spectra of BSA-Au NCs (0.1× dilution) in the presence of primary TB drugs (100 µM each in the final concentration). At 640 nm (emission wavelength): BSA-Au NCs (fluorescence intensity = 123.74 ± 1.56), NCs + rifampicin (83 µg/mL) (13.67 ± 0.33), NCs + pyrazinamide (12.3 µg/mL) (121.95 ± 0.69), NCs + ethambutol (27.7 µg/mL) (114.27 ± 1.28) and NCs + izoniazid (13.7 µg/mL) (127 ± 0.69) (each data point represents the average of three separate studies (n = 3), and the error bars denote the standard error of measurements within each experiment). b Comparison of the normalized decrease in fluorescence intensity of BSA-Au NCs (0.1× dilution) in the presence of primary TB drugs (100 µM each in final concentration). At 640 nm (emission wavelength): normalized quenching of NCs + rifampicin [(F0 − F)/F0 ~ 0.89 ± 0.006], NCs + pyrazinamide [0.013 ± 0.005], NCs + ethambutol [0.065 ± 0.008] and NCs + izoniazid [0.028 ± 0.007] (each data point represents the average of three separate studies (n = 3), and the error bars denote the standard error of measurements within each experiment). [In this work, (F0 − F)/F0 = 1 indicates complete quenching and (F0 − F)/F0 = 0 indicates no quenching]. The excitation wavelength was set at 480 nm, and the emission wavelength was 640 nm.
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Related In: Results  -  Collection

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Fig4: a Fluorescence emission spectra of BSA-Au NCs (0.1× dilution) in the presence of primary TB drugs (100 µM each in the final concentration). At 640 nm (emission wavelength): BSA-Au NCs (fluorescence intensity = 123.74 ± 1.56), NCs + rifampicin (83 µg/mL) (13.67 ± 0.33), NCs + pyrazinamide (12.3 µg/mL) (121.95 ± 0.69), NCs + ethambutol (27.7 µg/mL) (114.27 ± 1.28) and NCs + izoniazid (13.7 µg/mL) (127 ± 0.69) (each data point represents the average of three separate studies (n = 3), and the error bars denote the standard error of measurements within each experiment). b Comparison of the normalized decrease in fluorescence intensity of BSA-Au NCs (0.1× dilution) in the presence of primary TB drugs (100 µM each in final concentration). At 640 nm (emission wavelength): normalized quenching of NCs + rifampicin [(F0 − F)/F0 ~ 0.89 ± 0.006], NCs + pyrazinamide [0.013 ± 0.005], NCs + ethambutol [0.065 ± 0.008] and NCs + izoniazid [0.028 ± 0.007] (each data point represents the average of three separate studies (n = 3), and the error bars denote the standard error of measurements within each experiment). [In this work, (F0 − F)/F0 = 1 indicates complete quenching and (F0 − F)/F0 = 0 indicates no quenching]. The excitation wavelength was set at 480 nm, and the emission wavelength was 640 nm.
Mentions: We tested the selectivity of BSA-Au NCs for sensing rifampicin in the presence of other antibiotics (INZ, E and P) used collaterally with rifampicin for the treatment of TB. The fluorescent responses of BSA-Au NCs with all four antibiotics were monitored and compared with that of rifampicin. Figure 4a shows that the addition of rifampicin at a final concentration of 100 µM (83 µg/mL) resulted in a nearly 90% decrease in the fluorescence intensity of the BSA-Au NCs, whereas a limited decrease in fluorescence intensity was observed for P (1.3%), INZ (2.8%) and E (6.5%) at the same molar concentration (Figure 4b). The clinical effectiveness of rifampicin as a primary drug for TB can mainly be attributed to 4-methyl-1-piperazinaminyl substitution on rifampicin. Generally, antibiotics with piperazine moiety exhibit good efficacy as therapeutic agents [42–44]. Therefore, we tested the specificity of BSA-Au NCs toward detecting rifampicin using antibiotics with piperazine functional group such as ciprofloxacin [42], buspirone [43], ipsapirone [44] (Additional file 1: Figure S10a). In addition, we also measured the fluorescence responses of some commonly used antibiotics and drugs (Additional file 1: Figure S10b). The average amount of residual rifampicin present in the patients’ urine is in the range of 45–55 µg/mL [45]. Our results showed that at a final concentration of 50 µg/mL of rifampicin a nearly 60% decrease in the fluorescence intensity of the BSA-Au NCs was measured, whereas only a limited decrease in fluorescence intensity (less than 10%, p < 0.001) was observed for all the other antibiotics and drugs at the same concentration. Therefore, BSA-Au NCs can be used for preferential sensing of rifampicin among other antibiotics administered at the same time. Additionally, the prepared BSA-Au NCs were freeze-dried and stored for 3 months before dispersion for the same experiment (Additional file 1: Figure S11). No drastic changes were observed, even after storage, indicating the excellent stability of the BSA-Au NC.Figure 4

Bottom Line: The decreased fluorescence intensity of BSA-Au NCs in the presence of rifampicin allows for the sensitive detection of rifampicin in a range from 0.5 to 823 µg/mL.The detection limit for rifampicin was measured as 70 ng/mL.We have developed a robust, cost-effective, and portable point-of-care medical diagnostic platform for the detection of rifampicin in urine based on the ability of rifampicin to quench the fluorescence of immobilized BSA-Au NCs on wax-printed papers.

View Article: PubMed Central - PubMed

Affiliation: Department of Engineering and System Science, National Tsing Hua University, Hsinchu, 300, Taiwan. sanu.hit@gmail.com.

ABSTRACT

Background: Rifampicin or rifampin (R) is a common drug used to treat inactive meningitis, cholestatic pruritus and tuberculosis (TB), and it is generally prescribed for long-term administration under regulated dosages. Constant monitoring of rifampicin is important for controlling the side effects and preventing overdose caused by chronic medication. In this study, we present an easy to use, effective and less costly method for detecting residual rifampicin in urine samples using protein (bovine serum albumin, BSA)-stabilized gold nanoclusters (BSA-Au NCs) adsorbed on a paper substrate in which the concentration of rifampicin in urine can be detected via fluorescence quenching. The intensity of the colorimetric assay performed on the paper-based platforms can be easily captured using a digital camera and subsequently analyzed.

Results: The decreased fluorescence intensity of BSA-Au NCs in the presence of rifampicin allows for the sensitive detection of rifampicin in a range from 0.5 to 823 µg/mL. The detection limit for rifampicin was measured as 70 ng/mL. The BSA-Au NCs were immobilized on a wax-printed paper-based platform and used to conduct real-time monitoring of rifampicin in urine.

Conclusion: We have developed a robust, cost-effective, and portable point-of-care medical diagnostic platform for the detection of rifampicin in urine based on the ability of rifampicin to quench the fluorescence of immobilized BSA-Au NCs on wax-printed papers. The paper-based assay can be further used for the detection of other specific analytes via surface modification of the BSA in BSA-Au NCs and offers a useful tool for monitoring other diseases.

No MeSH data available.


Related in: MedlinePlus